1. Field of the Invention
This invention pertains to biological sensors and chemical detection methods, and more particularly to a protocol for conditioning honeybees to olfactory stimuli to aid biological sensor detection of agricultural plant pathogens. This invention also pertains to a memory enhancing cocktail that aids in long term memory retention of a vapor signature, thereby allowing the bees to be used in the field for longer durations and with fewer bees trained overall.
2. Description of Related Art
The presence of volatile and semi-volatile organic compounds (VOV/SVOC) can be a powerful diagnostic indicator in a variety of commercial applications including transportation security, search and rescue, narcotics interdiction, medical diagnostics, food quality and control, and facility security.
The detection of volatiles and semi-volatiles may be accomplished in the field with portable instruments, or in the laboratory with previously collected samples. Typically, chromatographic separations are performed by gas chromatography for identification. Some other technologies exist such as ion mobility spectrometry that can characterize some volatiles and semi-volatile organic compounds. These instruments and methods are both quantitative and qualitative in nature and can be costly to perform.
Non-instrument based detection methods in the art are primarily mammalian based systems. For example, canines are used extensively in smuggling interdiction, explosives detection, search and rescue, etc. where the plume of a volatile or semi-volatile set of chemicals is detected by a trained canine. Additionally, there are indicator papers that can change color when reacted with some analytes. The number of analytes that are detectable by this approach is very limited.
However, detection of volatile organic compounds in an agricultural setting can be complicated and many current methods may not be easily adapted to that setting. The agro-ecosystem is a complex biological arena, far from the sterile environment of a laboratory or the controlled environment of an international airport. Therefore, sensors for use in this system must be modified from that employed for sensors of explosives at national ports of entry, for example. Rather, a strong and ubiquitous background of volatile and semi-volatile organic compounds is present in a natural or agricultural setting, such as a commercial wine grape vineyard, that is often quite similar to the target volatile compound. For example, plants infected with pathogens or attacked by insects may release specific volatile organic compounds in response, but sometimes the volatile compounds that are indicative of such invasion may also be released from healthy, uninfected plants. The key difference may lie in amounts and/or ratios (to each other) of the volatile organic compounds that are released.
Accordingly, there is a need for inexpensive and accurate qualitative sensors for in situ detection of agricultural plant pathogens, thereby indicating disease. Such sensors will provide decision support for disease management, and potentially reduce the economic and environmental impact of remedial disease control measures. The present invention satisfies this need, as well as others, and is generally an improvement over sensors in the art.